Reproductive technology in horses encompasses a range of scientific techniques and procedures aimed at assisting and enhancing equine reproduction. These technologies include artificial insemination, embryo transfer, and cryopreservation of gametes and embryos. They are employed to improve breeding efficiency, manage genetic diversity, and preserve valuable genetic material. Artificial insemination involves the collection and introduction of semen into the mare's reproductive tract, while embryo transfer allows for the harvesting and implantation of embryos from donor to recipient mares. Cryopreservation involves freezing and storing sperm, oocytes, or embryos for future use. This page compiles peer-reviewed research studies and scholarly articles that investigate the methodologies, applications, and outcomes of reproductive technologies in equine breeding and management.
Hurtgen JP, Whitmore HL.Luteolysis, estrus, and ovulation were induced by endometrial biopsy during three periods of prolonged diestrus in two mares. Serum progesterone concentrations decreased to less than 1 ng/ml within 48 hours after biopsy. Mares were in estrus within 3 days after endometrial biopsy. It was concluded that endometrial biopsy may induce regression of the persistent corpus luteum by stimulating the release of an endogenous uterine luteolysin (prostaglandin).
Henneke DR, Kreider JL.Quarter Horse mares (n = 30) and their foals were used in a 2-year study. Objectives of the study were (i) to determine the effects of a treatment regimen, which permitted nursing for four 1-hour periods each day, on the postpartum reproductive performance of the dams, and (ii) to assess the effects of this procedure on the growth and development of foals. Mares in the restricted suckling treatment group showed estrus and ovulated sooner (P less than 0.01) after parturition than did control mares during the first year of the study, but not during the second year. Plasma progesterone concentrat...
Allen WE.The situations and conditions that can disrupt the mares normal oestrous cycle are described. Season of the year is a major influence; maximum reproductive efficiency does not totally coincide with the artificially defined "breeding season". Other abnormalities are associated with spontaneously persistent luteal function, psychological influences over behavioural activity, the presence of endometritis, multiple ovulation, reproductive behavior after pregnancy failure and granulosa cell tumours.
Thompson DL, Pickett BW, Squires EL, Amann RP.Factors affecting testicular measurements in situ and the relationships among the measurements and various reproductive characteristics were studied using data from 48 stallions. Mean values during the breeding season are provided for scrotal width, widths and lengths of individual testes, combined weight of testicular parenchyma, daily sperm production and daily sperm output. Testicular measurements were highly repeatable from day to day and for repeated measurements on a given day; technician provided the largest source of variation in the measurements of a given stallion. Age significantly ...
Betteridge KJ, Eaglesome MD, Flood PF.Two experiments were conducted using 14 mares. In Exp. 1, mares were inseminated with semen treated with TEPA, which, in other species, has been shown to lead to an arrest in ovum cleavage at 2--4 cells. The oviducts and/or uterus were then flushed 7--10 days after ovulation in 6 mares (Group A) or 2--6 days after ovulation in 5 mares (Group B). Fresh eggs were found in the oviduct flushes of 5 Group A and 5 Group B mares: 9 of the 10 eggs appeared to have cleaved, but none had developed beyond 16-cells. Seven eggs contained spermatozoa and 3 of 4 eggs from each group showed evidence of fertil...
Graham EF, Crabo BG, Pace MM.From the studies cited it was concluded that short and long term preservation of stallion semen has encountered major obstacles. Fertilizing capacity of extended or extended and cooled spermatozoa has been impaired. With the hydrogen ion extenders, the fertility was depressed either with or without glycerol when the semen was inseminated immediately after extension. With the cream-gel extender, fertility was not impaired when inseminated immediately after extension, but was impaired after storage at 5 C for 24 hr or in the presence of glycerol. The fertilizing capacity of extended frozen sperm...
Klug E, Brinkhoff D, Flüge A, Scherbarth R, Essich G, Kienzler M.Practical experiences of the phantom method for collection of genital secretions from stallions are reported. Taking a phantom used in the Richard-Götze-Haus Tierärztliche Hochschule Hannover as a prototype two further models slightly modified have been constructed, baring a flat hollow in the right side of the caudal phantom body for manual inserting of the Artificial Vagina. These three models fulfill four important conditions for routine use: (1) sufficient sexual attractivity for the stallions; 80-85% successful collections of presecretions out of a total of 1050 using the dummy and 70% ...
Hughes JP, Stabenfeldt GH.A brief review is presented of the use in equine veterinary medicine of anterior pituitary hormones, posterior pituitary hormones, placental hormones, steroid hormones and prostaglandins. These hormones are used frequently in clinical application without a scientific basis for their expected activity.
Thompson DL, Pickett BW, Berndtson WE, Voss JL, Mett TM.Stallions were subjected to a gradually increasing photoperiod beginning on October 15, 1973. The maximum artificial daylength (16 hr) was imposed on February 8, 1974, and maintained until October 6, 1974. Two ejaculates were collected from each of five treated and four control stallions weekly, with an interval of 24 hr between die paired ejaculates on the first and alternate weeks, and an interval of 1 hr on the second and alternate weeks. During summer, stallions subjected to the artificial photoperiod produced less (P<.05) gel-free semen and gel per ejaculate and had more (P<.05) spermatoz...
Merkt H.The use and techniques of artificial insemination for horses in Germany over the last 30 years is described. Artificial insemination appears to produce pregnancy percentages equal to those from normal breeding methods and its continued availability under veterinary supervision is recommended in conditions where disease, disability or distance debar normal service.
Tervit HR.Recent advances in reproductive physiology offer ways for exploiting superior, female cattle and for manipulating physiological events such as parturition. The techniques involved in these advances and their associated hazards are the subject of this review.
Allen WR, Bowen JM, Frank CJ, Jeffcott LB, Rossdale PD.This short review article describes the various techniques currently available for artificial insemination in the horse. The collection and use of raw and extended semen is discussed together with the more recent developments in freezing semen. The expected conception rates with both fresh and frozen semen are quoted. The possible benefits in disease control and stud management are discussed, as well as the difficulties in controlling the use of A.I. from the Breed Registration Authorities point of view.
Merkt H, Klug E, Krause D, Bader H.Stallion semen frozen by the pellet method showed no significant loss of sperm motility and fertility over long periods of storage in liquid nitrogen. Eighteen of thirty mares conceived after insemination with semen recovered in nine ejaculates from seven stallions and stored for 18 months to 7 years.
Azzie MA.The clinical effects and therapeutic value of a 500 mg implant of oestradiol benzoate was observed in thirty-nine Thoroughbred brood mares showing various abnormalities of the ovary and/or the reproductive tract. Sixteen similarly affected, untreated mares acted as controls. The implant was inserted during the early part of the normal anoestrous period and was removed 6 to 10 weeks later. The conception rate among the treated group (59%) during the onset of the ensuing breeding season was significantly higher (P less than 0-02) than that in the control group. The value of this form of treatmen...
Chandley AC, Fletcher J, Rossdale PD, Peace CK, Ricketts SW, McEnery RJ, Thorne JP, Short RV, Allen WR.Chromosomal abnormalities have been detected in seven mares isolated by their poor reproductive performance. All had small or rudimentary gonads and absent or irregular oestrous cycles. Two mares had an XO genotype, one was a 65,XXX female and another a 64,XY sex-reversed female. Two other mares were sex chromosome mosaics of the 63,X/64,XX type. The seventh mare showed a normal female karyotype but a small extra autosomal fragment was found in a few cells.
Pace MM, Sullivan JJ.Fertilization rate was highest in mares inseminated with frozen semen within 12 hr of ovulation. Foaling rate was improved (P less than 0-05) by increasing the number of motile spermatozoa inseminated from 40 X 10(6) to 80 X 10(6) but was not further improved by increasing the number to 160 X 10(6) or by increasing the frequency of insemination from once to twice daily. The fertilizing capacity of spermatozoa frozen in one of the hydrogen ion extenders studied was dependent upon relative osmotic pressure and method of freezing (ampoules or pellets). Adjusting glycerol concentration from 7% to ...
Guignot F, Ottogalli M, Yvon JM, Magistrini M.The objective of this study was to perform intracytoplasmic sperm injection (ICSI) on in vitro matured equine oocytes and to improve in vitro embryonic development on Vero cells after activation of the microinjected oocytes with calcium ionophore. After maturation (23 or 40 h, 38.5 degrees C, 5% CO2), the cumulus-oocyte complexes were denuded, centrifuged and all oocytes exhibiting the first polar body were microinjected. ICSI was performed using fresh semen from three fertile stallions. Microinjected oocytes were activated with calcium ionophore A23187 (10 min, 10 microM) and cultured individ...
Carnevale EM.Methods for the collection and transfer of equine oocytes have been developed, and uses of these techniques have resulted in new clinical and research possibilities. Because oocyte transfer avoids reproductive problems associated with the oviduct, uterus, and cervix, pregnancies can be produced from many mares that cannot carry a pregnancy or produce embryos. Oocytes for clinical transfers are usually collected from preovulatory follicles and cultured for a short interval or transferred directly into a recipient's oviduct. For oocyte transfer, the recipient is inseminated within the uterus. A ...
Taveiros AW, Oliveira MA, Lima PF, Tenório Filho F, Bartolomeu CC, Santos MH, Oliveira LR, Iunes-Souza TC, Freitas JC.Ten pluriparous mares were used as donors to supply embryos which were transferred into 103 recipients, 31 of which were nulliparous, 34 were pluriparous and lactating, and 38 were pluriparous and non-lactating. The embryos were recovered eight days after ovulation and pregnancy was confirmed by ultrasound six days after the transfer; the length of the embryos was measured ultrasonographically on days 12, 14, 16, 18, 20, 25 and 30 after the embryo transfer. One hundred and fifteen of 200 flushes provided embryos, 12 being degenerate and 103 being viable embryos. From the 103 embryo transfers c...
Sönmez C, Eroglu A.The present study was carried out to investigate the pregnancy rate after covering in the foal heat (Group I), in the subsequent spontaneous heat (Group II), and in the induced heat (Group III) after administration of 7.5 mg of the prostaglandin F2 alpha analogue Luprostiol (Reprodin, Bayer) between the 20th and 22nd day post partum. Breeding during foal heat resulted in a pregnancy rate of 43.9% compared to 48.6% in the subsequent spontaneous heat post partum. Of 18 mares in group III, 14 mares had a foal heat, whereas 4 others had not shown a foal heat. 1-8 days after treatment, 14 mares (77...
Coutinho da Silva MA, Carnevale EM, Maclellan LJ, Seidel GE, Squires EL.The objective of the study was to compare embryo development rates after transfer of oocytes collected 22 or 33 h after hCG injection into recipients inseminated within the uterus or the oviduct. Oocytes were collected at approximately 22 or 33 h after hCG injections and incubated for approximately 16 or 1.5 h, respectively, before transfer. Intrauterine inseminations using 1 x 10(9) progressively motile sperm were done approximately 12 h before and 2 h after transfer. For intraoviductal inseminations (gamete intrafallopian transfer [GIFT]), semen was centrifuged through a Percoll gradient, an...
Hisey EA, Ross PJ, Meyers S.As standard in vitro fertilization is not a viable technique in horses yet, many different techniques have been used to create equine embryos for research purposes. One such method is parthenogenesis in which an oocyte is induced to mature into an embryo-like state without the introduction of a spermatozoon, and thus they are not considered true embryos. Another method is somatic cell nuclear transfer (SCNT), in which a somatic cell nucleus from an extant horse is inserted into an enucleated oocyte, creating a genetic clone of the donor horse. Due to limited availability of equine oocytes in t...
Merlo B, Del Prete C, Mari G, Iacono E.The demand for equine in vitro produced embryos has increased over the last decade. The aim of this study was to compare the effects of an extended IVM or a prolonged period before fertilization, including holding time, on equine immature oocyte developmental competence. Oocytes, collected from abattoir-derived ovaries, were divided into 4 groups: H0/24 (n = 165) 0 h holding + standard 24-26 h IVM; H8/36 (n = 160) 8 h holding + 36 h IVM; H20/24 (n = 187) 20 h holding + 24 h IVM; H0/44 (n = 164) 0 h holding + 44 h IVM. Oocytes matured to MII were fertilized by intracytoplasm...
Ball BA, Brinsko SP, Thomas PG, Miller PG, Ellington JE.Development of 1- to 2-cell in vivo fertilized equine embryos cultured with or without uterine tubal epithelial cells (UTEC) was studied. One- to 2-cell embryos (n = 26) were collected surgically from the uterine tubes of pony mares 1 day after ovulation. Four- to 8-cell embryos (n = 9) were collected 2 days after ovulation. Presumptive zygotes and 2-cell embryos were cultured with (n = 17) or without (n = 9) UTEC, and all 4- to 8-cell embryos were cocultured with UTEC as positive controls. Uterine tubal epithelial cells were used as cell suspensions within 2 weeks after initiation of cultures...
Brand A, de Bois CH, Vandenhende R.Parenteral administration of 2.5-5 mg. of prostaglandin F2alpha to horses, 15 mg. to heifers or 25-30 mg to lactating cows and 15 mg. to sheep will induce regression of the corpus luteum (luteolysis) and a fertile oestrus within 48-72 hours. Because of their luteolytic effect prostaglandins may be used in various indications in the field of reproduction. An exception is the pig in which administration of prostaglandins does not induce luteolysis before D12 and therefore fails to induce oestrus. In horses, cattle and sheep, administration of prostaglandins during the first four days of the cycl...
Woods GL, Ginther OJ.The ability of a pituitary extract to induce ovulation and the effect of multiple ovulations on pregnancy rate and incidence of multiple fetuses were studied in seasonally anovulatory mares. The extract induced ovulation in 95% of mares having a greater than or equal to 25 mm follicle at the onset of treatment and in 64% the ovulations were multiple. The extract was less effective when the largest initial follicle was less than 25 mm. Approximately 50% of the treated mares that ovulated did not become pregnant or enter prolonged dioestrus before returning to an anovulatory condition. In treate...
Allen WE.The situations and conditions that can disrupt the mares normal oestrous cycle are described. Season of the year is a major influence; maximum reproductive efficiency does not totally coincide with the artificially defined "breeding season". Other abnormalities are associated with spontaneously persistent luteal function, psychological influences over behavioural activity, the presence of endometritis, multiple ovulation, reproductive behavior after pregnancy failure and granulosa cell tumours.
Rigoglio NN, Smith OE, Matias GSS, Miglino MA, Smith LC.Because of the growing importance of horses in leisure and several sports, somatic cell nuclear transfer (SCNT) is being used more frequently for cloning animals for performance and reproductive purposes. However, because of the need to perforate the zona pellucida during microsurgical reconstruction of the oocyte, it is possible that SCNT-derived embryos undergo premature hatching, resulting in embryo bisection and twinning. Therefore, because equine twin pregnancies often lead to abnormal embryo development and pregnancy failure, we performed a detailed comparative assessment of equine twin ...
Wehrenberg WB, Fowler S, Kurhajec J, Hutz RJ.Forty-four female ranch mink, maintained out-of-doors under standard conditions, were exposed to natural photoperiod supplemented with a period of artificial light from approximately 2300 hr to 0300 hr from early January to mid February. Breeding was initiated on March 1. After repeated attempts to breed the animals, it was determined that the likelihood of their breeding was very low, presumably due to the artificial and asynchronous long-day photoperiod. In an attempt to induce breeding, the mink were randomly assigned to one of four treatment groups: saline, 25 International Units (IU) PMSG...
Root Kustritz MV, Chenoweth PJ, Tibary A.To determine whether veterinarians perceive that theriogenology training at veterinary medical schools in North America and the Caribbean is adequate for achievement of theriogenology skills commonly used in private practice. Methods: A survey was mailed to members of the veterinary medical associations of Alabama, Kansas, Pennsylvania, and Washington. With regard to reproductive procedures in bovine, equine, porcine, small ruminant, camelid, and small animal species, veterinarians (predominantly practitioners) were asked to rate the importance of that procedure in their job and to assess thei...
Johnson AL.A study was conducted to evaluate the effectiveness of gonadotropin-releasing hormone (GnRH) pulse infusion to stimulate follicular development and induce ovulation in seasonally anestrous standardbred mares. Seventeen mares were selected for use in this experiment, on the basis of a previous normal reproductive history, and were housed under a photoperiod of 8L:16D beginning one week prior to the start of the experiment (second week in January). Mares were infused with 20 micrograms (n = 7) or 2 micrograms (n = 6) GnRH/h, or were subjected to photoperiod treatment only (controls, n = 4). Seru...
Allen WR, Bowen JM, Frank CJ, Jeffcott LB, Rossdale PD.This short review article describes the various techniques currently available for artificial insemination in the horse. The collection and use of raw and extended semen is discussed together with the more recent developments in freezing semen. The expected conception rates with both fresh and frozen semen are quoted. The possible benefits in disease control and stud management are discussed, as well as the difficulties in controlling the use of A.I. from the Breed Registration Authorities point of view.
Katila T.In this review, effects of the composition of the inseminate on uterine response and pregnancy rates in mares are discussed. The inseminate can differ for volume, sperm concentration, total sperm numbers, presence, absence, or proportion of seminal plasma, and extender composition. Semen can be used as fresh, cooled, or frozen. The site of semen deposition also plays a role; semen is deposited either into the uterine body (standard artificial insemination (AI)) or into the tip of the uterine horn ipsilateral to the preovulatory follicle (deep AI) using the hysterocopical or transrectally guide...
Witkowski M, Tischner M.The aim of the present study was to determine the effect of prolonged photoperiod during late pregnancy on subsequent ovarian activity and fertility in mares. Pregnant mares (n=13) due to give birth in January and February were stimulated by a fixed photoperiod (16 h light: 8 h dark) from 15 November (during the last 2-3 months of gestation) until up to 1 month after parturition. A control group of mares (n=9) due to give birth at the same time were kept in the same stable and management regimen, but under natural light conditions. Light-treated mares ovulated during foal oestrus approximately...
Binder H, Jakob C, Bucher P.Successful application of embryo transfer (ET) has become common practice in cattle, horses, sheep, goats and a variety of other species held in captivity. Yet in cattle only has the technique been established commercially. In 1994 more than 100,000 bovine embryos have been transferred in European countries. Important progress in transvaginal ovum pick up (OPU), in vitro production (IVP) and cryopreservation have further improved the applicability of ET. Direct transfer simplifies the procedure considerably allowing individual transfers and eliminating the need of synchronizing recipients. In ...
Ginther OJ, Bergfelt DR.Seasonally anovulatory mares were injected, i.m., twice daily with a GnRH analogue (GnRH-A), and hCG was given when the largest follicle reached 35 mm in diameter. In Exp. 1, treatment was initiated on 23 December when the largest follicle per mare was less than or equal to 17 mm. An ovulatory response (ovulation within 21 days) occurred in 17 of 30 (57%) GnRH-A-treated mares on a mean of 15.8 days. The shortest interval to ovulation in control mares (N = 10) was 57 days. The diameter of the largest follicle first increased significantly 6 days after start of treatment. In Exp. 2, treatment wa...
Cremonesi F, Anderson K, Lange-Consiglio A.TWO METHODS HAVE BEEN DESCRIBED TO RECOVER OOCYTES FROM EQUINE FOLLICLES IN EXCISED OVARIES: aspiration and scraping. Aim of this work was to develop an effective method for collecting equine oocytes using Tuohy needle and comparing this technique to aspiration and scraping, with or without tunica albuginea removal. This hollow hypodermic needle, usually employed for inserting epidural catheters, is designed with a slightly curved tip, shaped similar to a small curette. In unpeeled ovaries, the recovery rates of Tuohy needle group was higher (P < .05) than in the 16 g needle aspiration and ...
Bresnahan DR, Lupole RE, Stilz CR, Carnevale EM.Zona pellucida (ZP) proteins are important for fertilization and sperm binding and are closely associated with cumulus cells. Communication between cumulus and oocytes is facilitated by intracellular membrane channels composed of connexins. The extent aging impacts potential differences in fertilization and reductions in fertility is not well understood. This study characterized age-related differences in transcript abundance of ZP proteins and connexins in cells from ovarian follicles. Additionally, differences in sperm binding to oocytes from old and young mares was evaluated. For experiment...
Seidel GE.Principles for selecting future research projects include interests of investigators, fundability, potential applications, ethical considerations, being able to formulate testable hypotheses and choosing the best models, including selection of the most appropriate species. The following 10 areas of assisted reproduction seem especially appropriate for further research: efficacious capacitation of bovine spermatozoa in vitro; improved in vitro bovine oocyte maturation; decreasing variability and increasing efficacy of bovine superovulation; improved fertility of sexed semen; improving equine IV...
O'Donnell LJ, Sheerin BR, Hendry JM, Thatcher MJ, Thatcher WW, LeBlanc MM.The mare exhibits nocturnal uterine contractions in the last 6 days of gestation. It is hypothesized that estradiol 17beta (O17beta) may be associated with the nightly increase in uterine contractions. The 24-h secretion pattern of plasma O17beta was measured in 3 pony mares in late gestation to identify changes in release as the mare neared parturition. Blood was collected weekly at 08:00 hours beginning on day 240 and every third day from day 330 until delivery. Serial blood samples were collected from each mare every 30-min for 24-h beginning on gestation day 310 and every sixth day thereaf...
van Niekerk CH, Morgenthal JC, Sanders CP, Malan JE.Progesterone concentrations were assayed by a competitive protein-binding technique
in peripheral plasma samples collected twice
daily during four oestrous cycles of three
mares, and once a day during the first seven
weeks of pregnancy in four mares. Large
variations were found in progesterone levels
between morning and evening samples on the
same day in the same mare.
The lowest progesterone concentration
was found about the time of ovulation. Within 24 hours after ovulation the progesterone
concentration increased and two peaks, one
at 5 days and another at 8 days, were found.
Be...
Melchert M, Aurich J, Ertl R, Reichart U, Walter I, Gautier C, Kaps M, Aurich C.Mares resume ovarian activity rapidly after foaling. Besides follicle-stimulating hormone (FSH) and luteinizing hormone (LH), the pituitary synthesizes prolactin and growth hormone which stimulate insulin-like growth factor (IGF) synthesis in the liver. We tested the hypothesis that follicular growth is initiated already antepartum, mares with early and delayed ovulation differ in IGF-1 release and that there is an additional IGF-1 synthesis in the placenta. Plasma concentrations of LH, FSH, IGF-1, IGF-2, activin and prolactin. IGF-1, IGF-2, prolactin and their receptors in placental tissues w...
Teles Filho ACA, Sanchez DJD, Viana AGA, Sheheryar S, Guerreiro DD, Bustamante-Filho IC, Martins AMA, Sousa MV, Ricart CAO, Fontes W, Moura AA.The present study was conducted to investigate the global proteome of 8-day-old equine blastocysts. Follicular dynamics of eight adult mares were monitored by ultrasonography and inseminated 24 h after the detection of a preovulatory follicle. Four expanded blastocysts were recovered, pooled, and subjected to protein extraction and mass spectrometry. Protein identification was conducted based on four database searches (PEAKS, Proteome Discoverer software, SearchGUI software, and PepExplorer). Enrichment analysis was performed using g:Profiler, Panther, and String platforms. After the elimina...
Jung H, Yoon M.Spermatogonial stem cells originate from gonocytes and undergo self-renewal and differentiation to generate mature spermatozoa via spermatogenesis in the seminiferous tubules of the testis in male mammals. Owing to the unique capacity of these cells, the spermatogonial stem cell transplantation technique, which enables the restoration of male fertility by transfer of germlines between donor and recipient males, has been developed. Thus, spermatogonial stem cell transplantation can be used as an important next-generation reproductive and breeding tool in livestock production. However, in large ...
Lindinger H, Wehrend A.Various systems are available for birth monitoring in horses, whereby a distinction must be made between methods for more accurate prediction of the date of birth in order to intensify monitoring of the mare in a timely manner as well as methods for detecting individuals that are in labor. Basically, it should be noted that there are almost no studies that compare different methods on the same population of mares. As the time of birth approaches, physiological parameters of mare and fetus change, but their variability is too high to predict the exact parturition time point prospectively. The b...
Flores JG, Bueno VC, Bastos HBA, Rechsteiner SMDEF.Reproductive biotechniques in the equine species have advanced in the last decade and horse breeders have started to question the possibilities of interfering in the determination of foal sex. The aim of the present study was to verify whether the variables mares and stallion's age, side of the ovary containing the preovulatory follicle, preovulatory follicle diameter, time between breeding and ovulation, and ovulation inducing hormones influence the sex of the foal. A total of 259 reproductive cycles of 160 mares and 22 Thoroughbred stallions were used. Statistical analysis was performed usin...
